Team:KULeuven/Data

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<li class="off"><a href="https://2011.igem.org/Team:KULeuven/Project">Project</a>
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<li class="off"><a href="https://2011.igem.org/Team:KULeuven/Description">Project</a>
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<li><a href="https://2011.igem.org/Team:KULeuven/Project">Summary</a></li>
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<li><a href="https://2011.igem.org/Team:KULeuven/Description">Description</a></li>
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<li><a href="https://2011.igem.org/Team:KULeuven/Details">Extended</a><li>
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<li><a href="https://2011.igem.org/Team:KULeuven/Modeling">Modeling</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Modeling">Modeling</a></li>
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                <li><a href="https://2011.igem.org/Team:KULeuven/Thermodynamics">Thermodynamics</a></li>
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                <li><a href="https://2011.igem.org/Team:KULeuven/Applications">Applications</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Biobricks">Biobricks</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Biobricks">Biobricks</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Notebook">Notebook</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Notebook">Notebook</a></li>
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                <li><a href="https://2011.igem.org/Team:KULeuven/Results">Results</a></li>
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<li><a href="https://2011.igem.org/Team:KULeuven/Ethics">Ethics</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Ethics">Ethics</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Safety">Safety</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Safety">Safety</a></li>
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                <li><a href="https://2011.igem.org/Team:KULeuven/Law&Patents">Law&Patents</a></li>
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<h3>Data page</h3>
<h3>Data page</h3>
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<h2> How does <i>E.D. Frosti</i> work?</h2>
<h2> How does <i>E.D. Frosti</i> work?</h2>
<img src="http://homes.esat.kuleuven.be/~igemwiki/images/data/figure01.png"><br><br>
<img src="http://homes.esat.kuleuven.be/~igemwiki/images/data/figure01.png"><br><br>
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When Lactose is added to the medium, plasmid 1 will be induced to form INP via BBa_K584028.<br>
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The first illustration is a global overview of our project in which you can see how the plasmids (and the parts) function in our system. The next illustration shows the plasmids in more detail, with the favorite new parts and the characterized pre-existing parts. <br>
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When L-arabinose is added to the medium, plasmid 2 will be induced to form AFP via.
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<img src="http://homes.esat.kuleuven.be/~igemwiki/images/data/figure02.png"><br><br>
<img src="http://homes.esat.kuleuven.be/~igemwiki/images/data/figure02.png"><br><br>
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<b>Plasmid 1</b> contains the bricks necessary to induce expression of the red color (via the CrtEBI system) and  ice-nucleating protein (INP) in a lactose-dependent manner. For this plasmid, the INP has been cloned (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584024" target="blank"><b>BBa_K584024</b></a>) and tested for functionality by putting it behind the constitutive (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584027" target="blank"><b>BBa_K584027</b></a>) and lactose-inducible (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584028" target="blank"> <b>BBa_K584028</b> </a>) promoter.
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<br> <br>
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<b>Plasmid 2</b> contains the bricks that induce the expression of the black color (via the melA system) and  antifreeze protein (AFP) in a arabinose-dependent manner. For this plasmid, the AFP has been cloned (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584020" target="blank"> <b>BBa_K584020</b></a>) and tested for functionality by putting it behind the constitutive promoter (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584026" target="blank"><b>BBa_K584026</b></a>).<br><br>
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<b>Plasmid 3</b> contains the necessary bricks for our cell death mechanism. Briefly, lactose or arabinose will generate a ribolock-ceaB construct (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584015" target="blank" ><b> BBa_K584015</b></a> and <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584016" target="blank"> <b>BBa_K584016</b></a>).The Ribokey can be activated by a cold shock(<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584014" target="blank"> <b>BBa_K584014</b></a>). For this plasmid we cloned the lactose- and arabinose-inducible ceaB construct (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584016" target="blank"> <b>BBa_K584016</b></a> and <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584015" target="blank"> <b>BBa_K584015</b></a> resp.) and characterized the cold shock promoter using the Biobrick <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584004" target="blank"><b>BBa_K584004</b></a>.
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<img src="http://homes.esat.kuleuven.be/~igemwiki/images/data/figure03.png"><br><br>
<img src="http://homes.esat.kuleuven.be/~igemwiki/images/data/figure03.png"><br><br>
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All our obtained results can also be found at our <a href="https://2011.igem.org/Team:KULeuven/Results">results</a> section.
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<br><br>
<h2>Data for our favorite new parts</h2>
<h2>Data for our favorite new parts</h2>
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<li> 1. <b>BBa_K584028</b> <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584028" target="blank"> (Main Page) </a> - <b> pLac-Lux hybrid promoter + INP </b>, <b> BBa_K584028 </b> (K.U.Leuven, iGEM 2011): Tested with MilliQ water (first purified via reversed osmosis over 0,45 µm then filtered through a 0,22µm filter to remove all nucleating agents) at -6°C. We added 1mM IPTG to <i>E.D. Frosti</i>'s medium. Water freezes immediately upon addition. Addition of a control (GFP construct) <i> E.D. Frosti </i> did not result in freezing. <u>Note</u>: The promoter coupled to GFP did not show any leakiness, but when we coupled the promoter to the INP we noticed a certain leakiness. The dual inhibition system is a way to get around this leakiness.
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<li> 1. <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584024" target="blank"> <b>BBa_K584024</b> </a> - <b> INP extracellular generator </b> (K.U.Leuven, iGEM 2011): this biobrick generates INP that is expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584027" target="blank"><b>BBa_K584027</b></a>) and the lactose-inducible promoter (yielding  <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584028" target="blank"><b>BBa_K584028</b></a>).
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<li> 2. <b>BBa_K584027</b> <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584027" target="blank"> (Main Page) </a> - <b> Constitutive Promoter + INP </b>, <b> BBa_K584027 </b> (K.U.Leuven, iGEM 2011): Tested with MilliQ water (first purified via reversed osmosis over 0,45 µm then filtered through a 0,22µm filter to remove all nucleating agents) at -6°C. Water freezes immediately upon addition. Addition of a control (GFP construct) <i> E.D. Frosti </i> did not result in freezing.
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<li> 2. <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584020" target="blank"> <b>BBa_K584020</b> </a> - <b> AFP extracellular protein generator </b> (K.U.Leuven, iGEM 2011): this biobrick generates AFP that will be expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584026" target="blank"> <b>BBa_K584026</b></a>).
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<li> 3. <b>BBa_K584026</b> <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584026" target="blank"> (Main Page) </a> - <b> constitutive promoter + AFP </b>, <b> BBa_K584026 </b> (K.U.Leuven, iGEM 2011): We used the same conditions for the MilliQ water, but now at -10°C. Unfortunately addition of <i>E.D. Frosti</i> coated with AFP (or nothing: GFP control) did not result in prevention of ice formation. Due to lack of time we could not test other conditions (different temperatures, different concentrations,...)
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<h2> Data for pre-existing parts</h2>
<h2> Data for pre-existing parts</h2>
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<li> 1. <b>BBa_I13453</b> <a href="http://partsregistry.org/Part:BBa_I13453:Experience#User_Reviews" target="blank"> (Experience) </a> - <b> pBAD promoter</b>, <b> BBa_I13453 </b> (Endy Lab, iGEM 2005): Fused a GFP reporter to this promoter (which created biobrick: <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584000" target="blank"> <b>BBa_K584000</b></a>). The results can be found at the experience page of that promoter.
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<li> 1. <a href="http://partsregistry.org/Part:BBa_I13453:Experience#User_Reviews" target="blank"> <b>BBa_I13453</b> </a> - <b> pBAD promoter</b> (Endy Lab, iGEM 2005): fused a GFP reporter to this promoter (which created biobrick: <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584000" target="blank"> <b>BBa_K584000</b></a>). The results can be found at the experience page of the promoter.
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<li> 2. <b>BBa_J23119</b> <a href="http://partsregistry.org/Part:BBa_J23119:Experience" target="blank"> (Experience) </a> - <b> constitutive promoter</b>, <b> BBa_J23119 </b> (Berkeley, iGEM 2006): Fused a GFP reporter to this promoter (which created biobrick: <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584001" target="blank"> BBa_K584001</a>). The results can be found at the experience page of that promoter.
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<li> 2. <a href="http://partsregistry.org/Part:BBa_J23119:Experience" target="blank"><b>BBa_J23119</b> </a> - <b> constitutive promoter</b> (Berkeley, iGEM 2006): fused a GFP reporter to this promoter (which created biobrick: <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584001" target="blank"><b>BBa_K584001</b> </a>). The results can be found at the experience page of the promoter.
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<li> 3. <a href="http://partsregistry.org/Part:BBa_K091100:Experience" target="blank"><b>BBa_K091100</b> </a> - <b> Lac-Lux hybrid promoter</b> (Davidson-Missouri_Western, iGEM 2008): fused a GFP to this promoter (which created biobrick <a href="http://partsregistry.org/Part:BBa_K584002" target="blank"><b>BBa_K584002</b> </a>). The results can be found at the experience page of the promoter.
<h2>We've also characterized the following parts</h2>
<h2>We've also characterized the following parts</h2>
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<li> 1. <b>BBa_K584004</b> <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584004" target="blank"> (Main Page) </a> - <b> HybB promoter + GFP generator</b>, <b> BBa_K584004 </b> (KULeuven, iGEM 2011): We created this biobrick but due to lack of time we could not test it.
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<li> 1. <a href="http://partsregistry.org/Part:BBa_J45503:Experience" target="blank"> <b>BBa_J45503</b> </a> - <b> HybB promoter</b> (MIT, iGEM 2006): fused a GFP to this promoter (which yielded biobrick <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584004" target="blank"> <b>BBa_K584004</b> </a>). The results can be found at the experience page of the promoter.<br><br>
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<li> 2. <a href="http://partsregistry.org/Part:BBa_K091107:Experience" target="blank"> <b>BBa_K091107</b> </a> - <b> pLux/cI Hybrid Promoter</b> (Davidson-Missouri_Western, iGEM 2008): we wanted to use this promoter at the inception of our project, however on a 1% agarose gel we saw fragments of a different height than reported by previous iGEM competitions.<br><br>
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<li> 3. <a href="http://partsregistry.org/Part:BBa_K091104:Experience" target="blank"> <b>BBa_K091104</b> </a> - <b> pLac/Mnt Hybrid Promoter</b> (Davidson-Missouri_Western, iGEM 2008): we wanted to use this promoter at the inception of our project, however on a 1% agarose gel we saw fragments at a different height than reported by previous iGEM competitions.
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Latest revision as of 07:45, 28 October 2011

KULeuven iGEM 2011

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Data page

How does E.D. Frosti work?



The first illustration is a global overview of our project in which you can see how the plasmids (and the parts) function in our system. The next illustration shows the plasmids in more detail, with the favorite new parts and the characterized pre-existing parts.


Plasmid 1 contains the bricks necessary to induce expression of the red color (via the CrtEBI system) and ice-nucleating protein (INP) in a lactose-dependent manner. For this plasmid, the INP has been cloned (BBa_K584024) and tested for functionality by putting it behind the constitutive (BBa_K584027) and lactose-inducible ( BBa_K584028 ) promoter.

Plasmid 2 contains the bricks that induce the expression of the black color (via the melA system) and antifreeze protein (AFP) in a arabinose-dependent manner. For this plasmid, the AFP has been cloned ( BBa_K584020) and tested for functionality by putting it behind the constitutive promoter (BBa_K584026).

Plasmid 3 contains the necessary bricks for our cell death mechanism. Briefly, lactose or arabinose will generate a ribolock-ceaB construct ( BBa_K584015 and BBa_K584016).The Ribokey can be activated by a cold shock( BBa_K584014). For this plasmid we cloned the lactose- and arabinose-inducible ceaB construct ( BBa_K584016 and BBa_K584015 resp.) and characterized the cold shock promoter using the Biobrick BBa_K584004.


All our obtained results can also be found at our results section.

Data for our favorite new parts

  • 1. BBa_K584024 - INP extracellular generator (K.U.Leuven, iGEM 2011): this biobrick generates INP that is expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded BBa_K584027) and the lactose-inducible promoter (yielding BBa_K584028).

  • 2. BBa_K584020 - AFP extracellular protein generator (K.U.Leuven, iGEM 2011): this biobrick generates AFP that will be expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded BBa_K584026).

    Data for pre-existing parts

  • 1. BBa_I13453 - pBAD promoter (Endy Lab, iGEM 2005): fused a GFP reporter to this promoter (which created biobrick: BBa_K584000). The results can be found at the experience page of the promoter.

  • 2. BBa_J23119 - constitutive promoter (Berkeley, iGEM 2006): fused a GFP reporter to this promoter (which created biobrick: BBa_K584001 ). The results can be found at the experience page of the promoter.

  • 3. BBa_K091100 - Lac-Lux hybrid promoter (Davidson-Missouri_Western, iGEM 2008): fused a GFP to this promoter (which created biobrick BBa_K584002 ). The results can be found at the experience page of the promoter.

    We've also characterized the following parts

  • 1. BBa_J45503 - HybB promoter (MIT, iGEM 2006): fused a GFP to this promoter (which yielded biobrick BBa_K584004 ). The results can be found at the experience page of the promoter.

  • 2. BBa_K091107 - pLux/cI Hybrid Promoter (Davidson-Missouri_Western, iGEM 2008): we wanted to use this promoter at the inception of our project, however on a 1% agarose gel we saw fragments of a different height than reported by previous iGEM competitions.

  • 3. BBa_K091104 - pLac/Mnt Hybrid Promoter (Davidson-Missouri_Western, iGEM 2008): we wanted to use this promoter at the inception of our project, however on a 1% agarose gel we saw fragments at a different height than reported by previous iGEM competitions.